Objective To overcome the limitations of existing human respiratory syncytial virus(hRSV)animal models,such as semi-permissiveness and short duration of infection,this study established an IL2rg gene knockout(IL2rg-/-)rat model using TALEN gene editing technology.Methods The animal model was infected with hRSV intranasally.Clinical characteristics,body weight,and temperature changes were observed over the infection period(0～35 days).The total viral loads in respiratory organs,such as the nasal tissue,trachea,and lungs,were measured at various time points(4,11,20,and 35 days post-infection).Pathological analysis was conducted on target organs at the endpoint of observation(35 days post-infection).Changes in peripheral blood T,B,NK,and NKT cells and various cytokines were assessed at various time points(4,20,and 35 days post-infection).Results(1)IL2rg/-knockout rats sustained high viral loads in the nasal cavity upon intranasal inoculation with hRSV.The average peak titer rapidly reached 1 × 1010 copies/g in nasal tissue and 1 × 107 copies/g up to 5 weeks post-infection.(2)However,no significant pathological changes were noted in nasal,tracheal,or lung tissues.(3)An increase was observed in the content of peripheral blood B cells in hRSV-infected IL2rg--rats.(4)IL-6 and MCP-1 were increased in the early stage of infection and then decreased at the end of the observation period.Conclusions This study established a new IL2rg-/-rat model using TALEN technology and found that this model effectively supported high-level replication and long-term infection of hRSV,providing a good basis for antiviral drug screening and in vivo efficacy evaluation of anti-hRSV antibodies.

Alzheimer's disease(AD),the most common progressive neurodegenerative disease in dementia,is also a special lipid disease.From the perspective of modern medicine,cholesterol homeostasis is an important risk factor for AD.Amyloid-beta plaque deposition,neurofibrillary tangles,and large amount of lipid granule accumulation are typical pathological features of AD.From the perspective of TCM,turbidity is the key to the pathogenesis of AD.Phlegm turbid,stasis turbid and turbid toxin are the concrete derivation of turbidity,which are the standard of AD.Cholesterol is the greasy lipid which is produced from of the essence of water and food,the disturbance of cholesterol homeostasis is a typical embodiment of the pathogenesis mechanism of endogenous turbidity.Regulating cholesterol homeostasis by traditional Chinese medicine may be a new direction for the treatment of AD in the future.Focusing on the modern research of cholesterol homeostasis,taking the theory of turbidity as the starting point,this paper analyzed the correlation between the connotation of turbidity theory and the imbalance of cholesterol homeostasis as well as the pathogenesis mechanism,and further elucidated the clinical application results in the treatment of AD from the aspects of phlegm turbid,stasis turbid and turbid toxin,so as to better guide clinical practice and scientific research.

Objective: To investigate the prognostic utility of radiomics features extracted from 18 F-fluorodeoxyglucose (FDG) PET/CT combined with clinical factors and metabolic parameters in predicting progression-free survival (PFS) and overall survival (OS) in individuals diagnosed with extranodal nasal-type NK/T cell lymphoma (ENKTCL). Materials and Methods: A total of 126 adults with ENKTCL who underwent 18 F-FDG PET/CT examination before treatment were retrospectively included and randomly divided into training (n = 88) and validation cohorts (n = 38) at a ratio of 7:3.Least absolute shrinkage and selection operation Cox regression analysis was used to select the best radiomics features and calculate each patient’s radiomics scores (RadPFS and RadOS). Kaplan–Meier curve and Log-rank test were used to compare survival between patient groups risk-stratified by the radiomics scores. Various models to predict PFS and OS were constructed, including clinical, metabolic, clinical + metabolic, and clinical + metabolic + radiomics models. The discriminative ability of each model was evaluated using Harrell’s C index. The performance of each model in predicting PFS and OS for 1-, 3-, and 5-years was evaluated using the time-dependent receiver operating characteristic (ROC) curve. Results: Kaplan–Meier curve analysis demonstrated that the radiomics scores effectively identified high- and low-risk patients (all P < 0.05). Multivariable Cox analysis showed that the Ann Arbor stage, maximum standardized uptake value (SUVmax), and RadPFS were independent risk factors associated with PFS. Further, β2-microglobulin, Eastern Cooperative Oncology Group performance status score, SUVmax, and RadOS were independent risk factors for OS. The clinical + metabolic + radiomics model exhibited the greatest discriminative ability for both PFS (Harrell’s C-index: 0.805 in the validation cohort) and OS (Harrell’s C-index: 0.833 in the validation cohort). The time-dependent ROC analysis indicated that the clinical + metabolic + radiomics model had the best predictive performance. Conclusion The PET/CT-based clinical + metabolic + radiomics model can enhance prognostication among patients with ENKTCL and may be a non-invasive and efficient risk stratification tool for clinical practice.

Epidemiological and animal studies indicate that pre-existing diabetes increases the risk of Parkinson's disease(PD).However,the mechanisms underlying this association remain unclear.In the present study,we found that high glucose(HG)levels in the cerebrospinal fluid(CSF)of diabetic rats might enhance the effect of a subthreshold dose of the neurotoxin 6-hydroxydopamine(6-OHDA)on the development of motor disorders,and the damage to the nigrostriatal dopaminergic neuronal pathway.In vitro,HG promoted the 6-OHDA-induced apoptosis in PC12 cells differentiated to neurons with nerve growth factor(NGF)(NGF-PC12).Metabolomics showed that HG promoted hyperglycolysis in neurons and impaired tricarboxylic acid cycle(TCA cycle)activity,which was closely related to abnormal mito-chondrial fusion,thus resulting in mitochondrial loss.Interestingly,HG-induced upregulation of pyruvate kinase M2(PKM2)combined with 6-OHDA exposure not only mediated glycolysis but also promoted abnormal mitochondrial fusion by upregulating the expression of MFN2 in NGF-PC12 cells.In addition,we found that PKM2 knockdown rescued the abnormal mitochondrial fusion and cell apoptosis induced by HG+6-OHDA.Furthermore,we found that shikonin(SK),an inhibitor of PKM2,restored the mito-chondrial number,promoted TCA cycle activity,reversed hyperglycolysis,enhanced the tolerance of cultured neurons to 6-OHDA,and reduced the risk of PD in diabetic rats.Overall,our results indicate that diabetes promotes hyperglycolysis and abnormal mitochondrial fusion in neurons through the upre-gulation of PKM2,leading to an increase in the vulnerability of dopaminergic neurons to 6-OHDA.Thus,the inhibition of PKM2 and restoration of mitochondrial metabolic homeostasis/pathways may prevent the occurrence and development of diabetic PD.

Objective: To evaluate the immunity and influencing factors of diphtheria among preschool children in Shenzhen,to provide reference for effective monitoring of diphtheria IgG antibody level in preschool children. Methods: Serum samples were collected from 296 preschool children aged 4-6 who were recruited in Shenzhen. The diphtheria antibody titer in serum was determined by enzyme linked immunosorbent assay, and the effect of different immumuzation schedule including types of vaccine and vaccination timing, on the geometric mean concentration (GMC) of diphtheria IgG antibody and antibody positive rate were analyzed. Results: The GMC of diphtheria IgG antibody was 0.71 IU/mL, and the positive conversion rate was 33.1%. There were significant differences in antibody GMC and antibody positive conversion rate of diphtheria in different age groups( F/χ 2=11.77, 27.45, P < 0.01 ). The GMC and antibody positive conversion rate showed significant differences by diphtheria antibodies, vaccine types and end dose vaccination intervals( F=49.53, 12.95，11.61, P <0.01). There were statistically significant differences in the positive conversion rate of diphtheria antibodies in children with different types of diphtheria antibodies, vaccine types of diphtheria antibodies, and diphtheria antibodies at the time interval of final vaccination (Fisher exact probability method, P <0.01). Conclusion The overall positive conversion rate of diphtheria antibody in preschool children in Shenzhen is high. Timely completion of full diphtheria vaccination can improve the antibody level and plays a better role in protecting preschool children.

Objective:To evaluate rapid on-site evaluation (ROSE) in endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) for pancreatic solid lesions, and to compare the difference in ROSE performance between cytopathologists and trained endoscopists.Methods:A total of 168 consecutive patients with pancreatic solid lesions who underwent EUS-FNA from January 2014 to December 2020 at Fuding Hospital, Fujian University of Traditional Chinese Medicine were recruited. The patients who did not receive ROSE from January 2014 to November 2017 were included in N-ROSE group ( n=67). Since December 2017, the patients who intended to receive EUS-FNA were divided into E-ROSE group ( n=59, patients who received EUS-FNA and ROSE by endoscopists trained with cytopathology) and C-ROSE group ( n=42,patients who received EUS-FNA by untrained endoscopists and ROSE by cytopathologists) according to random number table. The number of punctures, sample adequacy, cytological diagnosis, final diagnosis and diagnostic efficiency (including the sensitivity, the specificity, the positive predictive value, the negative predictive value and the accuracy) in 3 groups were compared. Results:(1) The puncture number in N-ROSE group (4.22±0.76) was significantly more than E-ROSE group (3.12±0.79, P<0.001) and C-ROSE group (3.24±0.91, P<0.001). (2) The proportions of adequate samples in N-ROSE group [82.09% (55/67)] was significantly lower than those of E-ROSE group [96.61% (57/59), χ2=5.308, P=0.021] and C-ROSE group [97.62% (41/42), χ2=4.541, P=0.033]. The proportion of negative cytological diagnosis in N-ROSE group [40.30% (27/67)] was significantly higher than those of E-ROSE group [20.34% (12/59), χ2=5.848, P=0.016] and C-ROSE group [19.05% (8/42), χ2=5.348, P=0.021]. (3) The sensitivity of N-ROSE group [74.07% (40/54)] was significantly lower than those of E-ROSE group [94.00% (47/50), χ2=6.151, P=0.013] and C-ROSE group [94.44% (34/36), χ2=4.817, P=0.028]. The accuracy in N-ROSE group [79.10% (53/67)] was significantly lower than those of E-ROSE group [94.92% (56/59), χ2=5.433, P=0.020] and C-ROSE group [95.24% (40/42), χ2=4.155, P=0.042]. (4) There was no significant difference in any observational index between E-ROSE group and C-ROSE group ( P>0.05). Conclusion:ROSE in EUS-FNA can improve sample adequacy, the diagnostic sensitivity and accuracy, and reduce the number of punctures. The sample adequacy and diagnostic efficiency of endoscopists trained with cytopathology are comparable to those of cytopathologists.

Objective:To evaluate the value of transrectal ultrasound(TRUS)in diagnosing extramural venous invasion(EMVI) of rectal cancer.Methods:Clinical data of 81 rectal cancer patients were retrospectively analyzed in the People′s Hospital of Inner Mongolia Autonomous Region from January 2015 to December 2021. The extramural vascular sonographic features in these patients were summarized. Based on the postoperative pathology and compared with MRI examination, the efficacy of TRUS for the diagnosis of EMVI in rectal cancer was investigated.Results:①According to the sonographic presentation, extramural vessels of rectal cancer can be divided into the following four types: typeⅠshowed that the mass broke through the muscularis propria of the intestinal wall, and no peripheral vascular shadow was observed; typeⅡshowed that the mass broke through the muscularis propria, surrounded by extraneous vessels with normal diameter, running and blood flow; type Ⅲ showed that the mass broke through the intrinsic muscle layer of the intestinal wall and was surrounded by extramural vessels with abnormally thickened diameters, and blood flow filling defects; type Ⅳ showed a mass that broke through the intrinsic muscular layer, with irregularly dilated vessels outside the peritumoral wall and no blood flow signal in the lumen. ②With type Ⅲ and type Ⅳ as positive signs of EMVI, the diagnosis compliance rate of TRUS was 90.1% (Kappa=0.580, P<0.001), sensitivity was 58.3%, specificity was 95.7%, positive predictive value was 70.0%, negative predictive value was 93.0%, and area under the ROC curve(AUC) was 0.770. The diagnosis compliance rate of MRI was 86.4% (Kappa=0.541, P<0.001), sensitivity was 75.0%, specificity was 88.4%, positive predictive value was 52.9%, negative predictive value was 95.3%, and the AUC was 0.817. The differences in sensitivity and specificity between TRUS and MRI for the diagnosis of rectal cancer EMVI were not statistically significant ( P>0.05), and the differences in the AUC were not statistically significant ( Z=0.447, P=0.655). Conclusions:TRUS is valuable in assessing the extramural vascular status of rectal cancer and is expected to be an effective imaging method for preoperative diagnosis of EMVI.

Objective:To investigate the image characteristics of woven coronary artery (WCA)on intravascular ultrasound(IVUS) and optical coherence tomography(OCT).Methods:Thirty-seven patients suspected of WCA on coronary angiography were enrolled from Teaching Hospital of Chengdu University of Traditional Chinese Medcine, Zhengzhou Cardiovascular Disease Hospital and Zhongshan Hospital of Fudan University from January 2013 to July 2020. The intraluminal imaging features of WCA were analyzed using IVUS and OCT.Results:Of the 37 patients admitted at the cardiology service, 9 patients had WCA. All the patients underwent coronary angiography, IVUS and OCT, of which 6 lesions were located on the right coronary artery, 2 lesions were located on the left anterior descending artery and 1 patient had WCA on the circumflex artery. The mean length of WCA lesions was 2.2 cm(ranged from 1.2 cm to 4.5 cm). The angiographic appearance of WCA was numerous small tortious channels origined form the main lumen. The channels appeared to be " doughnut" like pattern and they merged to normal artery again after the anomalous segment. Flow limitation was rare unless there was coronary atherosclerosis. OCT and IVUS showed multiple spiral channels in the anomalous segment, which were independent of each other and each channels had a relatively complete three-layers vascular structure.Conclusions:With typical image characteristics, IVUS and OCT are able to screen out WCA and guide the treatment decision making.

Objective:To investigate the role of coupler perfusion in transrectal ultrasound in the diagnosis of preoperative T staging of rectal cancer.Methods:A retrospective analysis of the preoperative clinical data of 132 patients with rectal cancer in the People′s Hospital of Inner Mongolia Autonomous Region from June 2015 to November 2020. According to whether or not the patients agreed to coupler perfusion before ultrasound examination, they were divided into 2 groups, namely the perfusion group 69 cases and the non-perfusion group of 63 cases, with postoperative pathology as the gold standard, and compared with magnetic resonance imaging(MRI) to evaluate the accuracy of the 2 groups and MRI in the T staging of rectal cancer.Results:The total coincidence rates of the coupling agent perfusion group, non-perfusion group and MRI group for the diagnosis of rectal cancer T staging were 89.9%, 76.2% and 87.9%, respectively, and the difference among the three methods was statistically significant (χ 2=6.096, P=0.047). The diagnostic sensitivity of the coupling agent perfusion group for T1 stage was 96.0%, which was higher than 61.5% of the non-perfusion group and 92.3% of the MRI ( P=0.010). The specificity of the perfusion group for the diagnosis of T2 stage was 95.7%, higher than the non-perfusion group and MRI ( P=0.037), the positive predictive value of the perfusion group for T2 stage was 90.9%, which was higher than the non-perfusion group and MRI ( P=0.035). The diagnostic accuracy of the perfusion group for T2 stage was 94.2%, higher than the non-perfusion group and MRI (χ 2=7.070, P=0.029). There were no statistically significant differences in diagnostic sensitivity, specificity, positive predictive value, negative predictive value and accuracy among the perfusion group and the non-perfusion group and the MRI for T3 and T4 (all P>0.05). Conclusions:Coupled-agent perfusion makes it convenient and fast for intracavity ultrasound to diagnose T staging of rectal cancer, and the diagnostic efficiency is comparable to MRI. In particular, it can be used as a highly reliable imaging method for T1 and T2 rectal cancer.

Objective:To investigate the clinical application value of argon-helium knife cryoablation combined with radioactive seed implantation in the treatment of non-small cell lung cancer(NSCLC).Methods:A total of 117 patients with NSCLC admitted to Oncology Department of Fifth Affiliated Hospital of Zhengzhou University from January 2015 to January 2017 were included in our study.And they were divided into the combination group(n=63)treated with CT guided argon-helium knife cryoablation combined with radioactive 125I seeds implantation and the control group(n=54)treated only with argon-helium knife ablation.The changes of blood routine indexes, tumor markers, tumor ablation target volume and CT value were observed before and 1, 3, 6 months after treatment.Adverse reactions during treatment and the evaluation results of efficacy were compared between the two groups.Patients were followed up for 24 months to observe the recurrence and survival rates between the two groups. Results:In the combination group, seeds of(12.49±4.91)were implanted, and the X-ray exposure was(123.16±42.75)Gy.There was no significant difference in general clinical data between the two groups before treatment( P>0.05). At 1, 3 and 6 months after treatment, as compared with control group the combination group showed the significantly decreased platelet count( t=3.154, 3.586, 2.233, P=0.027、0.019、0.034), while, there was no significant difference in white blood cell count, red blood cell count and hemoglobin level between the two groups(all P>0.05). The levels of carcinoembryonic antigen(CEA), neuron-specific enolase(NSE)and tumor volume were significantly lower in combination group than in control group at 3 and 6 months after treatment( t3=3.142, 2.926 and 4.281, t6=4.094, 5.382 and 4.535, all P<0.05), showing significant improvements of illness.While, the above levels showed no significant differences at 1 month after treatment between two groups( t=1.065, 1.037, P=0.197, 0.255). At each monitoring time, the CT value of tumor target area showed a steady downward trend( P<0.05). During the treatment, the incidence of thrombocytopenia was higher in the combination group than in the control group(47.6% or 30/63 vs.24.1% or 13/54, χ2=6.935, P=0.008), while there were no significant differences in the incidence of postoperative fever, pneumothorax, myoglobinuria, pain, bleeding and nausea and vomiting between the two groups(all P>0.05). After 6 months of treatment, the remission rate was higher in the combination group(73.0% or 46/63)than in the control group(48.1% or 26/54). The survival time and relapse-free time of the combination group were longer than those of the control group[(21.81±4.31)months vs.(18.93±5.94)months, (20.48±5.76)months vs.(16.93±7.14)months, Log Rank χ2=8.229 and 9.656, P=0.004 and 0.002)]. Conclusions:Argon-helium knife Cryoablation combined with radioactive seed implantation can effectively control the local progression of NSCLC, reduce the risk of tumor recurrence, and has high safety.